近年來醫療的發達，科學家積極探索生理機制，而螢光探針便是不可或缺的存在。螢光探針具有方便、快速、專一性結合以及免於複雜前處理步驟等優點，隨著螢光探針的蓬勃發展，勢必面臨一些不可避免的缺點。像是在生物樣品中(如血漿)因為含有大量的蛋白質，而這些蛋白質會與螢光探針產生結合造成非專一性的訊號，導致螢光數據上的誤判而高估待測物質含量。而目前，螢光探針在生物樣品檢測中普遍擁有非專一性訊號的干擾，只能透過繁複的前處理作業才能達到偵測準確性，但對於生物研究上不夠方便快速。
　　在本研究中，我們開發一種新穎的螢光增益型探針可利用蛋白質作為盾牌，排除複雜環境中的蛋白質干擾，能以簡單快速且免於複雜前處理步驟的方式來偵測。本論文以分子轉子CCVJ和CCA為螢光基底，修飾上SNAP-tag假性受質O6-BG，也利用Biotin Avidin System來設計探針。當與蛋白質結合後，由於蛋白質巨大結構之立體障礙影響，使分子轉子單鍵旋轉的自由度下降，再利用硫化氫和硝基還原酶將辨識端移除，造成大幅度的螢光增益，也藉此達到偵測的效果。我們所設計的螢光探針具有合成容易、與目標蛋白質結合速率快且不需複雜樣品前處理等優勢。我們相信此種利用蛋白質保護螢光探針的設計可延伸運用於結合其他專一性蛋白與基團，或用於偵測不同的活性物質，並能在不受複雜蛋白質的影響下成功地運用在生物樣品中。

In recent years, small molecule fluorescent turn-on probes are getting much attention as they allow for sensitive, simple and specific detection with high signal-to-background ratios. Currently most of the small-molecule fluorescent turn-on probes are designed for monitoring proteins and reactive small molecules. Although most fluorescent probes show excellent performance in aqueous buffers, their application in protein rich human blood remain challenging as the real signal is usually obscured by nonspecific fluorescence.
Herein, we introduce a novel protein-shield concept to eliminate nonspecific fluorescence in protein rich blood samples. Based on this concept, we reveal that modified fluorescent probes conjugated to target protein can generate very specific fluorescence signals in blood serum by blocking out nonspecific binding of the dyes with other nonspecific blood proteins. By this protein-shield concept, we’ve successfully designed probe BTCCVJ- N3, BGCCA- N3 and BTCCVJ-NO2 to detect Nitroreductase (NTRase) and Hydrogen Sulfide (H2S) with fluorescence signal amplification and removal of nonspecific fluorescence signals.

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